PMID:23289425

Citation	Marti, R, Zurfluh, K, Hagens, S, Pianezzi, J, Klumpp, J and Loessner, MJ (2013) Long tail fibres of the novel broad-host-range T-even bacteriophage S16 specifically recognize Salmonella OmpC. Mol. Microbiol. 87:818-34
Abstract	We report isolation and characterization of the novel T4-like Salmonella bacteriophage vB_SenM-S16. S16 features a T-even morphology and a highly modified 160 kbp dsDNA genome with 36.9 mol % G+C, containing 269 putative coding sequences and three tRNA genes. S16 is a virulent phage, and exhibits a maximally broad host range within the genus Salmonella, but does not infect other bacteria. Synthesis of functional S16 full-length long tail fibre (LTF) in Escherichia coli was possible by coexpression of gp37 and gp38. Surface plasmon resonance analysis revealed nanomolar equilibrium affinity of the LTF to its receptor on Salmonella cells. We show that OmpC serves as primary binding ligand, and that S16 adsorption can be transferred to E. coli by substitution of ompC with the Salmonella homologue. S16 also infects 'rough' Salmonella strains which are defective in lipopolysaccharide synthesis and/or its carbohydrate substitution, indicating that this interaction does not require an intact LPS structure. Altogether, its virulent nature, broad host range and apparent lack of host DNA transduction render S16 highly suitable for biocontrol of Salmonella in foods and animal production. The S16 LTF represents a highly specific affinity reagent useful for cell decoration and labelling, as well as bacterial immobilization and separation.
Links	PubMed Online version:10.1111/mmi.12134
Keywords	Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Host Specificity; Host-Pathogen Interactions; Myoviridae/genetics; Myoviridae/metabolism; Porins/genetics; Porins/metabolism; Receptors, Virus/genetics; Receptors, Virus/metabolism; Salmonella Phages/genetics; Salmonella Phages/metabolism; Salmonella enterica/genetics; Salmonella enterica/metabolism; Salmonella enterica/virology; T-Phages/genetics; T-Phages/metabolism; Viral Tail Proteins/genetics; Viral Tail Proteins/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9CAUD:M1EAF4		GO:0098024: virus tail, fiber	ECO:0000314:		C		In silico analysis was used to identify gp38 as a component of the putative long tail fibre (LTF) (Figure 3). Sequence identity of S16 gp38 to the corresponding gene products in phages T4 and T2 is high (46%)(Figure 3). A phylogenetic comparison of gp38 of S16 to homologues of other T4-like phages also clusters the S16 variant closer to T2 than to T4 (Figure S2). SDS-PAGE of different S16 GFP-LTF protein synthesis variations in E.coli resulted in the production of functional GFP–LTF and indicates trimerization of S16 gp37 and attachment of gp38 to S16 gp37 trimers (Figure 4 and S1) The distal part of the S16 LTF likely has a putative structure closely related to what has been reported for phage T2 (Drexler et.al., 1986), where gp38 acts as the actual adhesin while bound to the C-terminal (distal) tip of gp37. The binding of purified S16 GFP–LTF fusion protein to wild-type and mutant Salmonella typhimurium cells (with mutated OmpC) were assessed. (Figure?5A and B) The wild-type revealed an even decoration of the bacterial surface by the fluorescent LTF (Figure?5A and B). Deletion of OmpC completely abolished decoration by S16 GFP–LTF (Fig.?5C and D). The defective phenotype could be restored by in-trans complementation using ompC (Table?3, Figure?5E and F).	complete CACAO 12625
9CAUD:M1EAL7		GO:0098024: virus tail, fiber	ECO:0000314:		C		In silico analysis was used to identify gp37 as a component of the putative long tail fibre (LTF)(Figure 3). Sequence identity of S16 gp37 to the corresponding gene products in phages T4 and T2 is very low (T4: 20% and T2: 18%). Because this homology is below the widely accepted threshold for relatedness (Tian and Skolnick, 2003), hence might be considered orthologues rather than homologues. SDS-PAGE of different S16 GFP-LTF protein synthesis variations in E.coli resulted in the production of functional GFP–LTF and indicates trimerization of S16 gp37 and attachment of gp38 to S16 gp37 trimers (Figure 4 and S1) The distal part of the S16 LTF likely has a putative structure closely related to what has been reported for phage T2 (Drexler et.al., 1986), where gp38 acts as the actual adhesin while bound to the C-terminal (distal) tip of gp37. The binding of purified S16 GFP–LTF fusion protein to wild-type and mutant Salmonella typhimurium cells (with mutated OmpC) were assessed. (Figure?5A and B) The wild-type revealed an even decoration of the bacterial surface by the fluorescent LTF (Figure?5A and B). Deletion of OmpC completely abolished decoration by S16 GFP–LTF (Figure?5C and D). The defective phenotype could be restored by in-trans complementation using ompC (Table?3, Figure?5E and F).	complete CACAO 12626

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